中国斑马鱼研究发展历程及现状

斑马鱼作为重要的脊椎动物模式系统之一, 由于其多方面的优势, 在生命科学研究领域发挥着越来越重要的作用。目前, 斑马鱼已被广泛地用于发育生物学、分子生物学、细胞生物学、遗传学、神经生物学、肿瘤学、免疫学、海洋生物学、药物学、毒理与环保等诸多方面的研究, 一些重要的成果不断涌现, 为现代生命科学的发展做出了重要贡献。我国20世纪90年代后期引入斑马鱼模式系统, 此后研究队伍扩大很快, 有影响的研究成果不断涌现, 促进了多个学科的发展。文章重点综述了我国内地与香港地区在斑马鱼研究方面的发展历程以及所取得的代表性成果, 以期促进该模式系统更广泛地用于开展高水平研究。     

斑马鱼血液疾病模型及应用

血液发育是一个复杂有序且保守的过程,由多种转录因子和信号通路协同调控,任何环节的失调都可能引起血液系统发育或功能缺陷,导致血液疾病的发生。斑马鱼(Daniorerio)造血过程及分子调控机制与哺乳动物高度保守。应用斑马鱼模拟致病因子的异常变化构建相关血液疾病模型,为探究疾病发生机制、肿瘤发生发展可视化研究及高通量化学筛选提供了有力的工具。本文概述了斑马鱼血液疾病模型及其应用,这些疾病模型不仅有助于完善对血液系统病理生理学、血液疾病发生分子机制的认识,也为临床相关恶性血液疾病的治疗提供了新思路。     

以斑马鱼为模式动物研究器官的发育与再生

斑马鱼因其受精卵体外发育、胚胎透明、具有较强的再生能力以及适于大规模遗传筛选的优势, 成为研究脊椎动物器官发育与再生的新兴模式动物。通过数十年的探索, 科研工作者已经在斑马鱼中建立了一套成熟的研究方法, 并对斑马鱼胚胎发育早期的细胞命运决定和分化、组织器官的形态建成以及受损后的再生过程有了初步的认识。近年来, 随着遗传筛选技术的大规模开展和活体成像技术在斑马鱼中的深入应用, 许多在小鼠等模式动物中悬而未决的问题开始得到充分解答。随着研究的不断深化和技术的不断更新, 以斑马鱼为模式动物, 对脊椎动物器官发育与再生的研究将会更加深入, 相关的调控机制也会被逐步探明, 从而为临床相关疾病的防治提供富有价值的参考。文章通过对近年来发表的文章进行回顾, 总结了斑马鱼作为模式动物研究中枢神经系统、肝脏和胰腺、血液细胞和血管等重要器官早期发育过程及其调控机制的进展, 并阐述了以斑马鱼研究尾鳍、心脏、肝脏等器官再生的优势和初步发现。     

斑马鱼生物钟研究进展

斑马鱼是生物钟研究领域中一种新兴的脊椎动物模型。文章总结了斑马鱼生物钟研究的一些进展, 以及利用斑马鱼研究生物钟的特点及优势。由于光照和温度作为重要的外部信号在斑马鱼生物钟调节中发挥重要作用, 文章主要就近期光和温度对斑马鱼钟基因及调节通路的研究进行了概述, 最后对斑马鱼生物钟研究的未来提出了展望。     

斑马鱼:在生命科学中畅游

模式动物是生命科学研究的重要材料,为生命科学的发展做出了重要的贡献。自20世纪90年代初以来,斑马鱼因其多方面的优点已成为模式动物家族中重要的一员,受到越来越多的重视和利用。目前,斑马鱼被广泛地用于发育生物学、遗传学、肿瘤学、药物学、毒理与环保等方面的研究,不断涌现新的研究成果。     

斑马鱼模型在药物筛选中的应用

斑马鱼具有子代数量多、体外受精、胚胎透明、可以做大规模遗传突变筛选等生物学特性, 因此成为一种良好的脊椎动物模式生物。随着研究的深入, 斑马鱼不仅应用于遗传学和发育生物学研究, 而且拓展和延伸到疾病模型和药物筛选领域。作为一种整体动物模型, 斑马鱼能够全面地检测评估化合物的活性和副作用, 实现高内涵筛选。近年来, 科学家们不断地发展出新的斑马鱼疾病模型和新的筛选技术, 并找到了一批活性化合物。这些化合物大多数在哺乳动物模型中也有相似的效果, 其中前列腺素E2(dmPGE2)和来氟米特(Leflunomide)已经进入临床实验, 分别用来促进脐带血细胞移植后的增殖和治疗黑素瘤。这些成果显示了斑马鱼模型很适合用于药物筛选。文章概括介绍了斑马鱼模型的特点和近年来在疾病模型和药物筛选方面的进展, 希望能够帮助人们了解斑马鱼在新药研发中的应用, 并开展基于斑马鱼模型的药物筛选。     

斑马鱼基因工程的研究进展

对国内外斑马鱼基因工程研究进展。包括最近获得的大批转基因斑马鱼品系、靶向筛选的转基因斑马鱼、斑马鱼转基因技术的发展和斑马鱼基因工程的热点问题与应用前景进行了综述。指出我国已建立日趋成熟的斑马鱼转基因技术、细胞核移植技术和染色体组操作技术,具有斑马鱼细胞遗传学和胚胎学基础,我国有望在不久的将来获得定向整合的基因工程斑马鱼,并推动生物技术应用研究领域的进步和发展。     

鱼类核移植与重编程

鱼类核移植是动物克隆研究的一个重要领域, 我国学者在上世纪60年代首创了鱼类的核移植研究。以斑马鱼为模式动物, 进行核移植与再程序化研究具有独特的优势。文章总结了鱼类细胞核移植研究的历史、斑马鱼核移植研究概况、以及影响核移植胚胎发育的因素, 特别是核移植胚胎基因组的表观遗传修饰, 如基因组DNA甲基化及组蛋白乙酰化和甲基化等的研究, 将有助于完善克隆技术并提高克隆的成功率, 推动克隆技术的广泛开展和应用。     

斑马鱼研究走向生物医学

1972年美国俄勒冈大学George Streisinger教授开始研究斑马鱼(Danio rerio)至今, 斑马鱼以其独特的优点, 已经成为现代遗传学、发育生物学研究的重要模式动物。世界范围内斑马鱼研究群体的工作已奠定了较为完善的胚胎学、分子遗传学研究基础, 并且斑马鱼已被应用于开发人类重大疾病模型和药物筛选平台, 取得了许多有价值的研究成果。文章简述了斑马鱼成为模式动物的历史, 侧重介绍了业已建立的白血病、黑色素瘤、感染免疫疾病、神经疾病等斑马鱼模型, 以及利用斑马鱼进行小分子化合物/药物筛选和研发的现状。斑马鱼研究向生物医学方向的拓展, 必将为人类理解重大疾病发生机制、寻找疾病治疗方法, 为维护人类卫生、健康做出贡献。     

斑马鱼,人类疾病研究的理想模式动物

斑马鱼作为一种理想的模式动物已有广泛的应用,而其基因组测序工程的完成和斑马鱼的基因与人类基因高度的相似性,使得斑马鱼在人类疾病的研究中体现着重要的价值.本文从斑马鱼在几种人类重大疾病研究中的运用的角度综述了斑马鱼作为一种重要的模式动物对人类疾病研究的贡献.     

Can zebrafish be used as a model to study the neurodevelopmental causes of autism?

The zebrafish has proven to be an excellent model for analyzing issues of vertebrate development. In this review we ask whether the zebrafish is a viable model for analyzing the neurodevelopmental causes of autism. In developing an answer to this question three topics are considered. First, the general attributes of zebrafish as a model are discussed, including low cost maintenance, rapid life cycle and the multitude of techniques available. These techniques include large-scale genetic screens, targeted loss and gain of function methods, and embryological assays. Second, we consider the conservation of zebrafish and mammalian brain development, structure and function. Third, we discuss the impressive use of zebrafish as a model for human disease, and suggest several strategies by which zebrafish could be used to dissect the genetic basis for autism. We conclude that the zebrafish system could be used to make important contributions to understanding autistic disorders.     

Zebrafish as a model for infectious disease and immune function

The zebrafish, Danio rerio, has come to the forefront of biomedical research as a powerful model for the study of development, neurobiology, and genetics of humans. In recent years, use of the zebrafish system has extended into studies in behaviour, immunology and toxicology, retaining the concept that it will serve as a model for human disease. As one of the most thoroughly studied teleosts, with a wealth of genetic and genomic information available, the zebrafish is now being considered as a model for pathogen studies in finfishes. Its genome is currently being sequenced and annotated, and gene microarrays and insertional mutants are commercially available. The use of gene-specific knockdown of translation through morpholino oligonucleotides is widespread. As a result, several laboratories have developed bacterial and viral disease models with the zebrafish to study immune responses to infection. Although many of the zebrafish pathogen models were developed to address human infectious disease, the results of these studies should provide important clues for the development of effective vaccines and prophylactic measures against bacterial and viral pathogens in economically important fishes. In this review, the capabilities and potential of the zebrafish model system will be discussed and an overview of information on zebrafish infectious disease models will be presented.     

Zebrafish heart as a model for human cardiac electrophysiology

The zebrafish (Danio rerio) has become a popular model for human cardiac diseases and pharmacology including cardiac arrhythmias and its electrophysiological basis. Notably, the phenotype of zebrafish cardiac action potential is similar to the human cardiac action potential in that both have a long plateau phase. Also the major inward and outward current systems are qualitatively similar in zebrafish and human hearts. However, there are also significant differences in ionic current composition between human and zebrafish hearts, and the molecular basis and pharmacological properties of human and zebrafish cardiac ionic currents differ in several ways. Cardiac ionic currents may be produced by non-orthologous genes in zebrafish and humans, and paralogous gene products of some ion channels are expressed in the zebrafish heart. More research on molecular basis of cardiac ion channels, and regulation and drug sensitivity of the cardiac ionic currents are needed to enable rational use of the zebrafish heart as an electrophysiological model for the human heart.     

Zebrafish kidney development: basic science to translational research

The zebrafish has become a significant model system for studying renal organogenesis and disease, as well as for the quest for new therapeutics, because of the structural and functional simplicity of the embryonic kidney. Inroads to the nature and disease states of kidney-related ciliopathies and acute kidney injury (AKI) have been advanced by zebrafish studies. This model organism has been instrumental in the analysis of mutant gene function for human disease with respect to ciliopathies. Additionally, in the AKI field, recent work in the zebrafish has identified a bona fide adult zebrafish renal progenitor (stem) cell that is required for neo-nephrogenesis, both during the normal lifespan and in response to renal injury. Taken together, these studies solidify the zebrafish as a successful model system for studying the broad spectrum of ciliopathies and AKI that affect millions of humans worldwide, and point to a very promising future of zebrafish drug discovery. The emphasis of this review will be on the role of the zebrafish as a model for human kidney-related ciliopathies and AKI, and how our understanding of these complex pathologies is being furthered by this tiny teleost.     

An investigation of the bioactivation potential and metabolism profile of Zebrafish versus human

The zebrafish model has been increasingly explored as an alternative model for toxicity screening of pharmaceutical drugs. However, little is understood about the bioactivation of drug to reactive metabolite and phase I and II metabolism of chemical in zebrafish as compared with human. The primary aim of our study was to establish the bioactivation potential of zebrafish using acetaminophen as a probe substrate. Our secondary aim was to perform metabolite profiling experiments on testosterone, a CYP3A probe substrate, in zebrafish and compare the metabolite profiles with that of human. The glutathione trapping assay of N-acetyl-p-benzoquinone imine demonstrated that zebrafish generates the same reactive metabolite as humans from the bioactivation of acetaminophen. Zebrafish possesses functional CYP3A4/5-like and UDP-glucuronosyltransferase metabolic activities on testosterone. Differential testosterone metabolism was observed among the two species. In silico docking studies suggested that the zebrafish CYP3A65 was responsible for the bioactivation of acetaminophen and phase I hydroxylation of testosterone. Our findings reinforce the need to further characterize the drug metabolism phenotype of zebrafish before the model can fully achieve its potential as an alternative toxicity screening model in drug research.     

FishNet: an online database of zebrafish anatomy

Background  

Over the last two decades, zebrafish have been established as a genetically versatile model system for investigating many different aspects of vertebrate developmental biology. With the credentials of zebrafish as a developmental model now well recognized, the emerging new opportunity is the wider application of zebrafish biology to aspects of human disease modelling. This rapidly increasing use of zebrafish as a model for human disease has necessarily generated interest in the anatomy of later developmental phases such as the larval, juvenile, and adult stages, during which many of the key aspects of organ morphogenesis and maturation take place. Anatomical resources and references that encompass these stages are non-existent in zebrafish and there is therefore an urgent need to understand how different organ systems and anatomical structures develop throughout the life of the fish.     

斑马鱼胚胎显微注射基因分析平台的建立

斑马鱼作为一种新的理想模式动物,已在发育生物学、环境毒理学和人类疾病及相关基因功能等研究中得到广泛应用。本文就建立斑马鱼胚胎的显微注射基因分析平台进行了探究,并通过piwil2基因的抑制和过表达实验验证了平台的可操作性,也对建立平台中所要注意的问题进行了讨论。     

Conservation of gene expression signatures between zebrafish and human liver tumors and tumor progression

The zebrafish (Danio rerio) has been long advocated as a model for cancer research, but little is known about the real molecular similarities between zebrafish and human tumors. Comparative analysis of microarray data from zebrafish liver tumors with those from four human tumor types revealed molecular conservation at various levels between fish and human tumors. This approach provides a useful strategy for identifying an expression signature that is strongly associated with a disease phenotype.     

The zebrafish fth1, slc3a2, men1, pc, fgf3 and cycd1 genes define two regions of conserved synteny between linkage group 7 and human chromosome 11q13

In addition to being an excellent model system for studying vertebrate development, the zebrafish has become a great tool for gene discovery by mutational analysis. The recent availability of the zebrafish EST database and radiation hybrid mapping panels has dramatically expanded the framework for genomic research in this species. Developing comparative maps of the zebrafish and human genomes is of particular importance for zebrafish mutagenesis studies in which human orthologs are sought for zebrafish genes. However, only partial cDNA sequences are determined routinely for mapped ESTs, leaving the identity of the EST in question. It previously had been reported that zebrafish linkage group 7 shares conserved synteny with human chromosome 11q13. In an effort to further define this relationship, five full-length zebrafish cDNAs, fth1, slc3a2, prkri, cd81, and pc, as well as one putative human gene, DBX were identified and their map positions ascertained. These six genes, along with men1, fgf3 and cycd1 define two regions of conserved synteny between linkage group 7 and 11q13.     

Zebrafish as a Model for Human Ciliopathies

Cilia, microtubule-based structures found on the surface of almost all vertebrate cells, play an array of diverse biological functions. Abnormal ciliary axonemal structure and function can result in a class of genetic disorders that are collectively termed ciliopathies. Model organisms,including Chlamydomonas reinhardtii and Caenorhabditis elegans have been widely used to study the complex genetic basis of ciliopathies.Here, we review the advantages of the zebrafish as a vertebrate model for human ciliopathies. We summarize the features of zebrafish cilia, and the major findings and contributions of the zebrafish model in recent studies of human ciliopathies. We also discuss the new genome editing approaches being efficiently used in zebrafish, and the exciting prospects of these approaches in modeling human ciliopathies.